Thrombin cleavage of GST from fusion proteins

Dyche Mullins 4/29/99

1. Elute the protein from the column using glutathione.

2. Add 10 units of thrombin per mg of protein.

3. Dialyze the protein + thrombin overnight at 4 degrees to remove glutathione.

4. Terminate the cleavage reaction with peptide inhibitor (ppack).

5. Reload mixture onto glutathione column to remove cleaved GST.

6. Clean up protein as necessary with additional chromatography steps.

Notes:

- Don't forget that GST is a dimer!

- Buy the good thrombin from Pharmacia

- ppack comes from Calbiochem